Evaluation of Compound Activity in Primary Human Intestinal Organoids Using Gene Expression and Histology

• DOI: 10.1002/cpph.54
• Publication Date: 2019-03-28T14:14:46Z
• Authors: Ronald Marchelletta; Jingxue Yu; Clara Moon; Mihee M. Kim
• Keywords: Organoid; Intestinal epithelium; Intestinal mucosa

Abstract Human intestinal organoids have enabled performance of functional epithelial studies and modeling human diseases the intestine. This unit describes 1) a method to isolate culture crypts from tissue, 2) use combinatorial methods expand stem cell–enriched spheroids differentiate them into composed various cell types, 3) stimulate these with measure their responsiveness external stimuli. To validate differentiation, can be stained qualitatively evaluate presence colonic crypt morphology specialized markers. These are responsive challenge tumor necrosis factor α (TNFα), resulting in cytokine‐induced apoptosis. TNFα‐driven apoptosis blocked by small‐molecule inhibitor Ire1α (4μ8C), an endoplasmic‐reticulum stress sensor. is one example how organoid model powerful tool elucidate important biological pathways involved disease cells. © 2019 John Wiley & Sons, Inc.